title: Detection of antibodies against SARS-Coronavirus using recombinant truncated nucleocapsid proteins by ELISA.
abstract: Severe acute respiratory syndrome (SARS) is a lifethreatening emerging respiratory disease caused by the coronavirus, SARS-CoV. The nucleocapsid (N) protein of SARS-CoV is highly antigenic and may be a suitable candidate for diagnostic applications. We constructed truncated recombinant N proteins (N1 [1-422 aa], N2 [1- 109 aa], and N3 [110-422 aa]) and determined their antigenicity by Western blotting using convalescent SARS serum. The recombinants containing N1 and N3 reacted with convalescent SARS serum in Western blotting. However, the recombinant with N2 did not. In ELISA using N1 or N3 as the antigens, positive results were observed in 10 of 10 (100%) SARS-CoV-positive human sera. None of 50 healthy sera gave positive results in either assay. These data indicate that the ELISA using N1 or N3 has high sensitivity and specificity. These results suggest that the middle or C-terminal region of the SARS N protein is important for eliciting antibodies against SARS-CoV during the immune response, and ELISA reactions using N1 or N3 may be a valuable tool for SARS diagnosis.
authors.affiliation: Department of Laboratory Animal Medicine, College of Veterinary Medicine, Seoul National University, Seoul 151-742, Korea.
authors.forename: Hyun-Kyoung
authors.initials: HK
authors.lastname: Lee
authors.forename: Byoung-Hee
authors.initials: BH
authors.lastname: Lee
authors.forename: Noton Kumar
authors.initials: NK
authors.lastname: Dutta
authors.forename: Seung-Hyeok
authors.initials: SH
authors.lastname: Seok
authors.forename: Min-Won
authors.initials: MW
authors.lastname: Baek
authors.forename: Hui-Young
authors.initials: HY
authors.lastname: Lee
authors.forename: Dong-Jae
authors.initials: DJ
authors.lastname: Kim
authors.forename: Yi-Rang
authors.initials: YR
authors.lastname: Na
authors.forename: Kyoung-Jin
authors.initials: KJ
authors.lastname: Noh
authors.forename: Sung-Hoon
authors.initials: SH
authors.lastname: Park
authors.forename: Hiroaki
authors.initials: H
authors.lastname: Kariwa
authors.forename: Mina
authors.initials: M
authors.lastname: Nakauchi
authors.forename: Le Quynh
authors.initials: le Q
authors.lastname: Mai
authors.forename: Suk-Jin
authors.initials: SJ
authors.lastname: Heo
authors.forename: Jae-Hak
authors.initials: JH
authors.lastname: Park
mesh_terms: D000914:Antibodies, Viral / Q000097:blood*; D000956:Antigens, Viral / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D004797:Enzyme-Linked Immunosorbent Assay / Q000379:methods*; D015870:Gene Expression; D006801:Humans; D019251:Nucleocapsid / Q000276:immunology* / Q000302:isolation & purification / Q000378:metabolism; D011994:Recombinant Proteins / Q000276:immunology / Q000302:isolation & purification / Q000378:metabolism; D045473:Severe acute respiratory syndrome-related coronavirus / Q000276:immunology*; D045169:Severe Acute Respiratory Syndrome / Q000175:diagnosis / Q000276:immunology*
chemical_list: D000914:Antibodies, Viral; D000956:Antigens, Viral; D011994:Recombinant Proteins